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Gene polymorphism detection kit for insulin resistance risk assessment and detection method and application of gene polymorphism detection kit

A technology for insulin resistance and gene polymorphism, which is used in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc. It can solve the problems of cumbersome and time-consuming operations, and simplify the sequencing process and time. Effect

Pending Publication Date: 2021-12-07
菲思特(上海)生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the methods of obtaining blood DNA mainly include traditional column extraction and magnetic bead extraction, both of which are time-consuming and cumbersome to operate

Method used

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  • Gene polymorphism detection kit for insulin resistance risk assessment and detection method and application of gene polymorphism detection kit
  • Gene polymorphism detection kit for insulin resistance risk assessment and detection method and application of gene polymorphism detection kit
  • Gene polymorphism detection kit for insulin resistance risk assessment and detection method and application of gene polymorphism detection kit

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Effect test

Embodiment 2

[0058] Embodiment 2, kit detection steps

[0059] The instruments adopted in the present invention are as follows: thermostat, pyrosequencer (Wuhan First Biotechnology Co., Ltd.).

[0060] 1) Take 30 μL EDTA anticoagulated whole blood sample in a PCR amplification tube;

[0061] 2) Add 100 μL of sample treatment solution and 4ul of magnetic beads, and let stand for 5 minutes;

[0062] 3) Place the PCR amplification tube on the magnetic stand, and suck out the mixed solution from the opposite side of the magnetic beads after all the magnetic beads are adsorbed to one side;

[0063] 4) Add amplification reagent 1 into the dry powder of amplification reagent 2, fully dissolve and mix;

[0064] 5) Add 25ul of the prepared PCR reaction solution into the PCR amplification tube obtained in step 4), fully mix the magnetic beads and the PCR reaction solution, centrifuge, and perform constant temperature reaction.

[0065] 6) Amplify using a PCR instrument, the reaction system is 25 ...

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Abstract

The invention discloses a gene polymorphism detection kit for insulin resistance risk assessment and a detection method and application of the gene polymorphism detection kit. The gene for the insulin resistance risk assessment is IRS-1, the kit designs a specific amplification primer and a sequencing primer aiming at the polymorphism of IRS-1 Gly972Arg, the kit comprises the following components: a sample treatment solution, magnetic beads, an amplification reagent 1, an amplification reagent 2, an IRS-1Gly972Arg sequencing primer and a positive control. According to the method, a biotin labeled single-stranded DNA is combined with streptavidin, the sequencing primer and a sequencing raw material are added after washing, pyrosequencing is carried out, the sequencing process is simplified, and the sequencing time is shortened. According to the kit, the gene polymorphism of the insulin resistance risk assessment is detected by combining rapid DNA preparation, constant-temperature PCR amplification and pyrosequencing technologies, and suggestions from the gene perspective are provided for clinical personalized medication.

Description

technical field [0001] The invention relates to a gene polymorphism detection kit for insulin resistance risk assessment and a detection method and application thereof, belonging to the field of gene detection. Background technique [0002] Insulin is a protein hormone secreted by pancreatic beta cells under the stimulation of endogenous or exogenous substances. Only when insulin binds to its receptor can it realize its physiological function in the body. Its main physiological function is to regulate the body's Sugar and fat metabolism, and promote protein synthesis. Insulin receptors are widely distributed in the body, and muscle, fat, liver and even kidney are all insulin target organs or tissues. Systemic or local insulin resistance (insulin resistance, IR) occurs when the sensitivity and responsiveness of insulin target organs or tissues to endogenous or exogenous insulin decrease. IRS1 (insulin receptor substrate 1), also known as IRS-1, is the main substrate of insu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12Q1/6869C12Q1/6844C12N15/11
CPCC12Q1/6883C12Q1/6869C12Q1/6844C12Q2600/156C12Q2600/106C12Q2565/301C12Q2521/507C12Q2522/101C12Q2531/119C12Q2563/143C12Q2563/149
Inventor 陈立波孙悦刘丹
Owner 菲思特(上海)生物科技有限公司
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